Multiplexed array of nanoliter droplet array device

ABSTRACT

A device comprising: plurality of Stationary Nanoliter Droplet Array (SNDA) components; each SNDA component comprising: at least one primary channel; at least one secondary channel; and a plurality of nano-wells that are each open to the primary channel and are each connected by one or more vents to the secondary channel; the vents are configured to enable passage of air solely from the nano-wells to the secondary channel, such that when a liquid is introduced into the primary channel it fills the nano-wells, and the originally accommodated air is evacuated via the vents and the secondary channel/s; an inlet port and a distribution channel configured to enable a simultaneous introduction of the liquid into all primary channels; and an outlet port and an evacuation channel configured to enable a simultaneous evacuation of the air out of all the secondary channels.

FIELD OF THE INVENTION

The present invention relates to microfluidic devices. Moreparticularly, the present invention relates to a multiplexed array ofnanoliter droplet array devices.

BACKGROUND OF THE INVENTION

Microfluidic devices that are designed to hold nanoliter-sized dropletsof liquids in separate nano-wells, referred to herein as a stationarynanoliter droplet array (SNDA) devices, have been proven to be of use inthe execution of various biological and chemical tests and procedures.In a typical procedure, two or more fluids are introduced successivelyinto the device via one or more inlets. The nano-wells are thenexamined, e.g., visually by: a microscope, an automated image analysissystem, or other visualization tools, to determine results of anyinteractions between the successively introduced liquids, or effects oncells that are suspended in one of the introduced liquids.

In a typical SNDA device, the introduced fluid may flow from the inletinto a primary channel of the device. The primary channel is lined onboth sides by openings to nano-wells, where adjacent nano-wells arebeing separated one from another by walls. An end of each nano-well thatis distal to its opening to the primary channel includes one or morevents that are opened to an air evacuation channel. Thus, as eachnano-well is filled with liquid via its opening to the primary channel,air that had previously filled the nano-well escapes through its vent tothe air evacuation channel. The openings of the vent are typically smallenough so as to prevent the liquid from passing out of the nano-wellthrough the vent. For example, the liquid may be prevented from emergingthrough the vent by the action of surface tension, viscosity, airpressure, or other forces. Thus, each nano-well may be partially orcompletely filled by the introduced liquid.

For example, such SNDA devices have been employed successfully toperform antimicrobial susceptibility testing (AST). When an SNDA deviceis used for AST, an antibiotic liquid is first introduced into each ofthe nano-wells. In some cases, the antibiotic may be introduced into thenano-wells in a manner that produces a gradient of concentration of theantibiotic along the length of the primary channel. The antibiotic maybe lyophilized or otherwise treated, e.g., to retain the antibiotic inthe nano-wells. A bacterial suspension may then be introduced into thenano-wells. The nano-wells may then be examined to determine the effectof the antibiotic on the bacteria. For example, an image of the SNDAdevice may be analyzed, either by a human eye or by a processor, todetermine the effect of the antibiotic on the bacteria.

SUMMARY OF THE INVENTION

According to some embodiments of the invention, a new device is providedcomprising:

-   -   plurality of Stationary Nanoliter Droplet Array (SNDA)        components; each SNDA component comprising: at least one primary        channel; at least one secondary channel; and a plurality of        nano-wells that are each open to the primary channel and are        each connected by one or more vents to the secondary channel;        the vents are configured to enable passage of air solely from        the nano-wells to the secondary channel, such that when a liquid        is introduced into the primary channel it fills the nano-wells,        and the originally accommodated air is evacuated via the vents        and the secondary channel/s;        -   wherein the plurality of the SNDA components are aligned            parallel to one another and laterally displaced relative to            one another, such that the device comprises a rectangular            form;    -   an inlet port and a distribution channel configured to enable a        simultaneous introduction of the liquid into all primary        channels; and    -   an outlet port and an evacuation channel configured to enable a        simultaneous evacuation of the air out of all the secondary        channels.

According to some embodiments, the diameter D_(DCh), or the smaller sideh_(DCh) of the distribution channel, is selected to be substantiallylarger than the diameter D_(PCh), or the smaller side h_(PCh) of theprimary channel/s, respectively D_(DCh)>D_(PCh) or h_(DCh)>h_(PCh); suchthat the distribution channel is configured to be filled via the inletport with liquid, while withholding the liquid from the primarychannels, to about a predetermined threshold of its volume, enabling aliquid pressure, formed there-within, to then simultaneously load allthe primary channels.

According to some embodiments, the device further comprising a pluralityof distribution channels, each distribution channel of the plurality ofdistribution channels connecting the inlet port to the primary channelof a separate SNDA component; and wherein each distribution channelbranches off from a single trunk channel that is connected to the inlet.

According to some embodiments, each distribution channel branches offperpendicularly from the trunk channel.

According to some embodiments, each of the distribution channelscomprises a different cross section, relative to its distance from thecommon inlet, configured to allow a liquid flow from the common inletopening, to flow via the common distribution channel, and reach all ofthe SNDA components concurrently.

According to some embodiments, the distribution channels are arrangedalong the trunk channel symmetrically, about a connection of the inletto the trunk channel.

According to some embodiments, the connections the plurality ofdistribution channels with the trunk channel are equally spaced alongthe trunk channel.

According to some embodiments, a total length of each of eachdistribution channel of the plurality of distribution channels, betweenits connection to the trunk channel and its connection to the primarychannel of an SNDA component, is adjusted to enable the substantiallyequal rates of liquid flow.

According to some embodiments, the total length of at least onedistribution channel of the plurality of distribution channels islengthened by addition of one or more open loops to said at least onedistribution channel.

According to some embodiments, the lengths of all of the open loops thatare added to distribution channels of the plurality of distributionchannels are substantially equal.

According to some embodiments, the length of an open loop of said one ormore open loops is equal to a distance between connections of twoadjacent distribution channels of the plurality of distribution channelsto the trunk channel, where the connections of the plurality ofdistribution channels to the trunk channel are equally spaced along thetrunk channel.

According to some embodiments, the number of the open loops that areadded to a first distribution channel is smaller than the number of theopen loops that are added to a second distribution channel, wherein aconnection of the second distribution channel to the trunk channel ismore proximal to a connection of the inlet to the trunk channel than tothe connection of the first distribution channel to the trunk channel.

According to some embodiments, a cross section of a distributionchannel, of the plurality of distribution channels, is selected toenable the substantially equal rates of liquid flow into each of theprimary channels.

According to some embodiments, a width of a distribution channel havinga largest cross-sectional area, is equal to a width of the primarychannel of the SNDA component to which that distribution channel isconnected.

According to some embodiments, all of the SNDA components aresubstantially identical.

According to some embodiments, the device further comprising a pressuredevice in communication with the outlet poet, configured to applysimultaneous negative pressure to all the secondary channels via theevacuation channel.

According to some embodiments of the present invention, there isprovided an array of stationary nanoliter droplet array (SNDA) devices.The array may include a plurality of the SNDA devices aligned parallelto one another and laterally displaced relative to one another, eachSNDA device of the plurality of SNDA devices comprising a primarychannel and a plurality of nano-wells that are each open to the primarychannel, each nano-well of said plurality of nano-wells being connectedby one or more vents to a secondary channel to enable passage of airfrom that nano-well to the secondary channel when a liquid that isintroduced into the primary channel fills that nano-well.

The array may also include an inlet for enabling introduction of theliquid into the array; and a plurality of distribution channels, eachdistribution channel of the plurality of distribution channelsconnecting the inlet to the primary channel of a separate SNDA device ofthe plurality of SNDA devices.

In some embodiments of the present invention, each distribution channelof the plurality of distribution channels branches off from a singletrunk channel that is connected to the inlet.

In some embodiments of the invention, each distribution channel of theplurality of distribution channels branches off perpendicularly from thetrunk channel.

In some embodiments of the invention, the plurality of distributionchannels are arranged along the trunk channel symmetrically about aconnection of the inlet to the trunk channel.

In some embodiments of the invention, connections the plurality ofdistribution channels with the trunk channel are equally spaced alongthe trunk channel.

In some embodiments of the invention, a total length of each of eachdistribution channel of the plurality of distribution channels betweenits connection to the trunk channel and its connection to the primarychannel of an SNDA device of the plurality of SNDA devices is adjustedto enable the substantially equal rates of flow.

In some embodiments of the invention, the total length of at least onedistribution channel of the plurality of distribution channels islengthened by addition of one or more open loops to said at least onedistribution channel.

In some embodiments of the invention, the lengths of all of the openloops that are added to distribution channels of the plurality ofdistribution channels are substantially equal.

In some embodiments of the invention, the length of an open loop of saidone or more open loops is equal to a distance between connections of twoadjacent distribution channels of the plurality of distribution channelsto the trunk channel where the connections of the plurality ofdistribution channels to the trunk channel are equally spaced along thetrunk channel.

In some embodiments of the invention, the number of the open loops thatare added to a first distribution channel is smaller than the number ofthe open loops that are added to a second distribution channel, whereina connection of the second distribution channel to the trunk channel ismore proximal to a connection of the inlet to the trunk channel than tothe connection of the first distribution channel to the trunk channel.

In some embodiments of the invention, a cross section of a distributionchannel of the plurality of distribution channels is adjusted to enablethe substantially equal rates of flow.

In some embodiments of the present invention, a width of a distributionchannel of the plurality of distribution channels having a greatestcross-sectional area is equal to a width of the primary channel of theSNDA device of the plurality of SNDA devices to which that distributionchannel is connected.

In some embodiments of the invention, all of the plurality of SNDAdevices are substantially identical.

In some embodiments of the invention, the secondary channels of theplurality of SNDA devices are connected to a single evacuation channel.

In some embodiments of the invention, the evacuation channel or sharedchannel includes an opening via which negative pressure is applicable toall of the secondary channels of the plurality of SNDA devices.

In some embodiments of the invention, plurality of the arrays is eachconnected to a single input opening via a feeder channel, all of thefeeder channels configured to enable concurrent loading of the arrays.

In some embodiments of the invention, all arrays of the plurality ofarrays are oriented parallel to one another.

In some embodiments of the invention, the feeder channels are branched.

In some embodiments of the invention, an array of the plurality ofarrays is oriented perpendicular to at least one other array of theplurality of arrays.

In some embodiments of the invention, the secondary channels of theplurality of SNDA devices of all arrays of the plurality of arrays areconnected to a single evacuation channel or shared secondary channel.

In some embodiments of the invention, the evacuation channel or sharedchannel includes an opening via which negative pressure is applicable toall of the secondary channels of the plurality of SNDA devices of allarrays of the plurality of arrays.

BRIEF DESCRIPTION OF THE DRAWINGS

The subject matter regarded as the invention is particularly pointed outand distinctly claimed in the concluding portion of the specification.The invention, however, both as to organization and method of operation,together with objects, features, and advantages thereof, may best beunderstood by reference to the following detailed description when readwith the accompanying drawings in which:

FIG. 1A schematically illustrates an example of a plurality ofstationary nanoliter droplet array (SNDA) components arranged in anarray configuration, forming a rectangular multiplexed SNDA device,according to some embodiments of the invention;

FIG. 1B schematically illustrates another example of a rectangularmultiplexed SNDA device, according to some embodiments of the invention;

FIG. 1C schematically illustrates yet another example of a plurality ofSNDA components arranged in an array configuration, forming arectangular multiplexed SNDA device, according to some embodiments ofthe invention;

FIG. 2 schematically illustrates an arrangement of distribution channelsof a portion of a multiplexed array of SNDA devices, according to someembodiments of the invention;

FIG. 3 schematically illustrates distribution channels a multiplexedarray of SNDA devices, the lengths of the channels being adjusted andconfigured to enable a uniform flow rate, according to some embodimentsof the invention;

FIG. 4A schematically illustrates an example of channels of a system ofmultiple multiplexed SNDA device arrays, where all SNDA devices areoriented parallel to one another, according to some embodiments of theinvention; and

FIG. 4B schematically illustrates an example of channels of a system ofmultiple multiplexed SNDA device arrays, where some SNDA devices areoriented perpendicularly to others, according to some embodiments of theinvention.

It will be appreciated that for simplicity and clarity of illustration,elements shown in the figures have not necessarily been drawn to scale.For example, the dimensions of some of the elements may be exaggeratedrelative to other elements for clarity. Further, where consideredappropriate, reference numerals may be repeated among the figures toindicate corresponding or analogous elements.

DETAILED DESCRIPTION OF THE INVENTION

In the following detailed description, numerous specific details are setforth in order to provide a thorough understanding of the invention.However, it will be understood by those of ordinary skill in the artthat the invention may be practiced without these specific details. Inother instances, well-known methods, procedures, components, modules,units and/or circuits have not been described in detail so as not toobscure the invention.

Although embodiments of the invention are not limited in this regard,discussions utilizing terms such as, for example, “processing,”“computing,” “calculating,” “determining,” “establishing”, “analyzing”,“checking”, or the like, may refer to operation(s) and/or process(es) ofa computer, a computing platform, a computing system, or otherelectronic computing device, that manipulates and/or transforms datarepresented as physical (e.g., electronic) quantities within thecomputer's registers and/or memories into other data similarlyrepresented as physical quantities within the computer's registersand/or memories or other information non-transitory storage medium(e.g., a memory) that may store instructions to perform operationsand/or processes. Although embodiments of the invention are not limitedin this regard, the terms “plurality” and “a plurality” as used hereinmay include, for example, “multiple” or “two or more”. The terms“plurality” or “a plurality” may be used throughout the specification todescribe two or more components, devices, elements, units, parameters,or the like. Unless explicitly stated, the method embodiments describedherein are not constrained to a particular order or sequence.Additionally, some of the described method embodiments or elementsthereof can occur or be performed simultaneously, at the same point intime, or concurrently. Unless otherwise indicated, the conjunction “or”as used herein is to be understood as inclusive (any or all of thestated options).

In accordance with some embodiments of the invention, and asdemonstrated in FIGS. 1A, 1B and 1C, plurality of stationary nanoliterdroplet array (SNDA) components 14 (twelve SNDA components, in theseexamples) are arranged in an array configuration, forming a rectangularmultiplexed SNDA device 10. In the multiplexed SNDA device 10, a liquidmay be introduced into the nano-wells 18 of all of the SNDA components14 of the SNDA device 10 via a common inlet opening 12, also referred toherein as a shared inlet. From the common inlet opening 12, theintroduced liquid flows through an arrangement of distribution channels46 that connects the inlet opening to the primary channel 16 of eachSNDA component 14. As the liquid flows along the primary channel of eachSNDA component, the liquid fills the nano-wells 18 along that primarychannel.

According to some embodiments, in the multiplexed SNDA device 10, aplurality of SNDA components 14 are arranged substantially parallel toone another and are substantially aligned with one another. In thisparallel and aligned configuration, the primary channels 16 of the SNDAcomponents 14 are parallel to one another and are laterally displacedrelative to one another. Thus, in this configuration, the connections ofall of the primary channels to distribution channel/s lie along a singleline 34, e.g., a line that is perpendicular to the orientation of theprimary channels.

According to some embodiments, as the liquid fills the nano-wells 18 ofeach SNDA component 14, air escapes via vent/s (not shown/visible) ofeach nano-well 18 into a secondary channel/s 20. According to someembodiments, each SNDA component 14 typically includes two secondarychannels 20, configured such that air from the nano-wells on either sideof the primary channel 16 is enabled to vent out of the nano-well 18.According to some embodiments, in the multiplexed SNDA device 10, all ofthe secondary channels are arranged to connect to a single evacuationchannel 22. According to some embodiments, concurrently withintroduction of liquid, via the common inlet opening 12, negativepressure can be applied to the evacuation channel 22, via an outlet 44,configured to facilitate removal of the air from the nano-wells, and tofacilitate flow of the introduced liquid into the nano-wells.

According to some embodiments, the distribution channels are configuredsuch that a liquid that is introduced via the common inlet opening 12flows into each primary channel 16 of the SNDA components 14 of themultiplexed SNDA device 10, at substantially equal flow rates. Forexample, flow rates may be considered to be substantially equal, whenthe differences in flow rate between two distribution channels does notexceed 5%, or, in some cases, does not exceed 3%. In this manner, thenano-wells of all of the SNDA components 14, in the multiplexed SNDAdevice 10, fill concurrently and at a common flow rate.

According to some embodiments, and as specifically demonstrated in FIG.1A, some SNDA components 14, of the multiplexed SNDA device 10, arecloser to the common inlet opening 12 than others. Therefore, accordingto some embodiments, a wide distribution channel 25 is provided, as aconnecting channel between the single inlet 12 and the primary channels16, feeding the wells 18 of the SNDA components 14. Accordingly, thecross-section of the wide distribution channel 25 is selected to belarger than the cross-section of the primary channels, such that thewide distribution channel 25 is configured to be filled with liquid, toa predetermined level of it's volume, before the liquid pressure that isformed there-within enables the liquid to flow and enter into theprimary channel/s 16. According to some embodiments, the cross-sectionof the distribution channel 25 and/or the primary channel/s comprises aform selected from: a circle, an oval, a rectangle, a square, anypolygon and any combination thereof.

According to some embodiments, the cross-section of the distributionchannel 25 and the primary channel/s comprises a circular form.Accordingly the diameter D_(DCh) of the wide distribution channel 25 isselected to be larger than the diameter D_(PCh) of the primary channel/s16 (D_(DCh)>D_(PCh)), such that the wide distribution channel 25 isconfigured to be filled with liquid to a predetermined threshold (for anon-limiting example about 95%-99%) of its volume, before the liquidpressure that is formed there-within enables to liquid to enter into theprimary channel/s 16, in other words, before the liquid pressure that isformed there-within raises high enough, to enable the liquid to flowagainst the primary channel/s flow resistance.

According to some related embodiments, where their cross section iscircular, an important solution to the Navier-Stokes equations is thePoiseuille (or Hagen-Poiseuille) flow, which applies when a pressuregradient is used to drive a liquid through a capillary or channel. For acapillary with cylindrical cross-section the following expression forthe volume flow, Q, exists:

$\begin{matrix}{Q = {\frac{\Delta V}{t} = {\frac{\pi R^{4}V}{8\eta L}\Delta P}}} & {{Eq}.\mspace{14mu}\left\{ 1 \right\}}\end{matrix}$

where R is the radius of the capillary, L is its length and ΔP is thepressure drop across this length (also called hydraulic pressure). Theterm, 8ηL/πR⁴, of which the reciprocal appears in Eq. {1}, is alsocalled the fluidic resistance. The dependency on 1/R⁴ implies that thefluidic resistance increases drastically as the channel dimensions arereduced. Consequently, higher pressure drops are necessary to moveliquid through smaller conduits. For channels with noncylindrical crosssections, expressions similar to those in Eq. {1} can be found, but withdifferent terms for the fluidic resistance.

According to some related embodiments, where their cross section iscircular, the ratio between D_(DCh):D_(PCh) is respectively selectedfrom: 10:1, 9:1, 8:1, 7:1, 6:1, 5:1 and any combination thereof.According to some embodiments, the ratio between D_(DCh):D_(PCh) isrespectively 4 or more:1. According to some embodiments, the ratiobetween D_(DCh):D_(PCh) is respectively selected X:1 where X is selectedbetween: 10>X>4.

According to some embodiments, the cross-section of the distributionchannel 25 and the primary channel/s comprises a rectangular form. Forthis example, shown in FIG. 1A, AA is the cross section of the widedistribution channel, where h_(DCh) is the smaller side and w_(DCh) isthe other side of the AA rectangular cross section and BB is the crosssection of the primary channel, where h_(PCh) is the smaller side andw_(PCh) is the other side of the BB rectangular cross section. Accordingto such embodiments, the wall dimension h_(DCh) of the wide distributionchannel 25 is selected to be larger than the wall dimension h_(PCh) ofthe primary channel/s 16 (h_(DCh)>h_(PCh)), such that the widedistribution channel 25 is configured to be filled with liquid to apredetermined threshold (for a non-limiting example about 95%-99%) ofits volume, before the liquid pressure that is formed there-withinenables to liquid to enter into the primary channel/s 16, in otherwords, before the liquid pressure that is formed there-within raiseshigh enough, to enable the liquid to flow against the primary channel/sresistance. According to a non-limiting example: AA=w_(DCh)×h_(DCh)=0.3mm×0.3 mm and BB=w_(PCh)×h_(PCh)=0.15 mm×0.1 mm.

According to some related embodiments, where their cross section isrectangular, an important solution to the Navier-Stokes equations is thePoiseuille (or Hagen-Poiseuille) flow, which applies when a pressuregradient is used to drive a liquid through a capillary or channel. For acapillary with rectangular cross-section the following expressionapproximation for the volume flow, Q, exists:

$\begin{matrix}{Q = {\frac{\Delta V}{t} = {\frac{h^{4}}{12\eta La}\Delta{P\left( {1 - {{0.6}3a}} \right)}}}} & {{Eq}.\mspace{14mu}\left\{ 2 \right\}}\end{matrix}$

where h is the smaller wall, and w is the other wall of the capillary, Lis its length, a=h/w is the aspect ratio of capillary walls, and ΔP isthe pressure drop across this length (also called hydraulic pressure).The term, 12ηLa/h⁴, of which the reciprocal appears in Eq. {2}, is alsocalled the fluidic resistance. The dependency on 1/h⁴ implies that thefluidic resistance increases drastically as the channel dimensions arereduced. Consequently, higher pressure drops are necessary to moveliquid through smaller conduits.

According to some related embodiments, where their cross section isrectangular, the ratio between h_(DCh):h_(PCh) is respectively selectedfrom: 10:1, 9:1, 8:1, 7:1, 6:1, 5:1 and any combination thereof.According to some embodiments, the ratio between h_(DCh):h_(PCh) isrespectively 4 or more:1. According to some embodiments, the ratiobetween h_(DCh):h_(PCh) is respectively selected X:1 where X is selectedbetween: 10>X>4.

According to some embodiments, and as demonstrated in FIGS. 1B, 1C, 2,3, 4A and 4B, some SNDA components 14 of the multiplexed SNDA device 10are nearer/closer to the common inlet opening 12 than others. Therefore,a distribution channel 24 f,27 f that connects the common inlet opening12 to a nearer SNDA component is configured to resist, or introduce adelay, into the flow through that distribution channel, relative to adistribution channel 24 a,27 a that connect a more distant SNDAcomponent to the common inlet opening.

According to some embodiments, the distribution channel/s 24 thatconnect the common inlet opening 12 with the SNDA components that areclose/r to the inlet opening are configured to be lengthened by anaddition of bends or open loops 24 b, 24 c, 24 d, 24 e, 24 f. In thismanner, the lengths of all distribution channels 24 a, 24 b, 24 c, 24 d,24 e, 24 f that connect each SNDA component to the common inlet openingare equal. For example, where the flow through the distribution channelsis assumed to be laminar, and where all of the distribution channelshave substantially identical cross sections, the resistance to flow isassumed to be simply proportional to the length of the channel. In thiscase, where the flow rate is assumed to be equal to the pressuredifference divided by the resistance to flow (analogous to Ohm's law forelectrical current, potential difference, and electrical resistance,respectively), a calculation of the required additional length that isto be added to each distribution channel to ensure identical flow ratesmay be similar to analogous calculations for simple electrical circuitsbased on Kirchhoff's rules for electrical circuits.

According to some embodiments, alternatively or in addition, thecross-sectional area of a shorter distribution channel, e.g., thatconnects the common inlet opening to a nearer SNDA device is configuredwith a narrower diameter than a longer distribution channels thatconnects the common inlet opening to a more distant SNDA component.

According to some embodiments, and as specifically demonstrated in FIG.1C, a flow resistance to the liquid entering from common inlet 12 via acommon distribution channel 28 is configured to be made significantlylow at a distal distribution channels of a distal SNDA component 14 a(for example 27a is distal from inlet 12), compared with a proximaldistribution channels of a proximal SNDA component 14 f (for example 27fis proximal to inlet 12), such that flow rate entering to each of theprimary channels is about equal. According to some embodiments, areduced cross-sectional area is configured to reduce a flow rate,through a proximal distribution channel, relative to the flow ratethrough a distal distribution channel. In this way, the liquid thatflows through the distribution channels 27 from the common inlet opening12, via the common distribution channel 28, reaches all of the SNDAcomponents 14 concurrently.

In some related embodiments, the connecting channels are designeddifferently one from another (by length as in 24 FIG. 1B, or by width asin 27 FIG. 1C) and/or that the resistance at the common distributionchannel (as in 25 FIG. 1A) is configured to be reduced, such that fluidcan first fill the common distribution channel 25,28 and then flowthrough the SNDAs' main channels to enable simultaneous loading of theSNDA components.

According to some embodiments, in addition to the introduction of aliquid into all of the SNDA components 14 of the multiplexed SNDA device10, via the common inlet opening 12, the primary channel of each SNDAdevice can include an individual opening 32, configured to enableselective introduction of liquid into selected individual SNDAcomponents 14. Typically (but not necessarily), the individual openingof each primary channel is located at an end of the primary channel thatis opposite the opening of the primary channel to the distributionchannels. For example, different experiments can be conductedconcurrently, by introducing different antibiotic solutions, or thatreagent solutions can be introduced into different SNDA component.According to some embodiments, no antibiotic or reagent solutions shouldbe introduced into an SNDA component that is to function as a controlmeasure.

According to some embodiments, the multiplexed SNDA device 10 comprisesa flat rectangular form, such that all SNDA components 14 are arrangedin an array configuration and are oriented parallel to one another andlinearly displaced relative to one another along a single pair oforthogonal axes. This rectangular arrangement within the multiplexedSNDA device 10 is advantageous over other arrangements of SNDA devices(e.g., a circular arrangement, where SNDA devices extend radially froman inlet opening). For example, the rectangular arrangement isconfigured to enable more efficient use of space/volume, e.g., morecompact filling, than an arrangement where adjacent SNDA components arerotated relative to one another. The rectangular arrangement isconfigured to enable efficient and easy control of the SNDA components,for example when positioning (whether manually or by an automaticallycontrolled stage) a successive SNDA component within a field of view ofa viewing or imaging device.

According to some embodiments, a plurality of rectangular multiplexedSNDA devices 10 are configured to be connected to a common inlet, asdemonstrated in FIGS. 4A and 4B. For example, the plurality ofrectangular multiplexed SNDA devices 10 can be connected to the commoninlet in a symmetric manner such that the lengths of channels thatconnect the common inlet to the inlet opening of each of the multiplexedSNDA device 10 are equal to one another. In some cases, one or more ofthe multiplexed SNDA device arrays can be rotated 90° relative to otherof the multiplexed SNDA device. When one multiplexed SNDA device isrotated by 90° relative to another, the aforementioned advantages ofefficient use of space and ease of control may still be present.

Reference is made again to FIG. 1B, which schematically illustrates anexample of a rectangular multiplexed array 10 of stationary nanoliterdroplet array (SNDA) components 14, according to some embodiments of theinvention.

In some embodiments, the multiplexed SNDA device 10 is provided aplurality of SNDA components 14, which are arranged parallel to oneanother. A liquid may be introduced concurrently into all SNDAcomponents 14 via common inlet 12, also referred to herein as sharedinlet 12. For example, common inlet 12 may connect to an opening in acover (not shown) that covers multiplexed SNDA device 10.

According to some embodiments, the common inlet 12 is connected to eachof the SNDA components 14 via a distribution channel 24. In the exampleshown, distribution channels 24 branch off of a single distributiontrunk channel 28. According to some embodiments, and as in the shownexample, distribution channels 24 branch off perpendicularly fromdistribution trunk channel 28. In other examples/embodiments,distribution channels 24 can otherwise connect to common inlet 12. Forexample, a distribution channel 24 can connect to common inlet 12 via adiagonal or curved segment of that distribution channel 24, can branchoff of distribution trunk channel 28 at an oblique angle, or mayotherwise connect to common inlet 12.

According to some embodiments, and as in the shown example, common inlet12 is located at symmetry axis 30, and distribution channels 24 arearranged symmetrically about symmetry axis 30. In otherexamples/embodiments, common inlet 12 can be located closer to onelateral side of multiplexed SNDA device array 10, e.g., such that adistance between common inlet 12 and an SNDA component 14 at one end ofmultiplexed SNDA device 10 is less than the distance between commoninlet 12 and an SNDA component 14 at the other end of multiplexed SNDAdevice 10.

According to some embodiments, each SNDA component 14 comprises aprimary channel 16 that connects to one of distribution channels 24.Thus, a liquid that is introduced into common inlet 12 can flow fromcommon inlet 12 and into primary channels 16 of all SNDA components 14of multiplexed SNDA device 10 via distribution channels 24 that connectcommon inlet 12 to all primary channels 16.

According to some embodiments, a separate inlet 32 (located at anopening in a cover of multiplexed SNDA device 10) to each primarychannel 16 can be located at an end of primary channel 16 that isopposite to an end that is connected via distribution channel 24 tocommon inlet 12. Accordingly, liquid can be introduced into primarychannel 16 of a selected SNDA components 14 of the multiplexed SNDAdevice 10 via separate inlets 32 of the selected SNDA components 14,without being introduced into other SNDA components 14 of themultiplexed SNDA device array 10.

According to some embodiments, a liquid that flows into a primarychannel 16 of an SNDA component 14 can flow into nano-wells 18 that areopen to that primary channel 16. As each nano-well 18 is filled, any airor gas that had previously filled that nano-well 18 is enabled to flowoutward via one or more vents of that nano-well 18 (not visible at thescale of FIG. 1B) to a secondary channel 20 that is adjacent to thatnano-well 18. For example, a typical SNDA component 14 includes twosecondary channels 20, on opposite sides of its primary channel 16.

In some embodiments, each nano-well 18 typically has a volume that isless than 100 nanoliters. In some embodiments, each vent has a length ofa few (less than or about 10) μm. In some embodiments, each nano-well 18has a length about 400 μm, a width of about 200 μm, and a height ofabout 100 μm, each vent has a width of about 7 μm and a height of about100 μm, each primary channel 16 (and, possibly, each distributionchannel 24) has a width of about 150 μm, and each secondary channel 20has a width of about 1 mm. In other examples, structure of a multiplexedSNDA device 10 can have different dimensions.

In the example shown, all secondary channels 20 of multiplexed SNDAdevice 10 connect to a single evacuation channel 22. In this manner, airfrom all nano-wells 18 can be evacuated via a single opening 44.According to some embodiments, negative pressure that is applied toevacuation channel 22 is, therefore, applied to all secondary channels20 and to all nano-wells 18. Thus, application of negative pressure toevacuation channel 22 facilitates flow of liquid into nano-wells 18.

According to some embodiments, the structure of multiplexed SNDA device10, including channels (e.g., common inlet 12, distribution trunkchannel 28, distribution channels 24, primary channels 16, separateinlets 32, secondary channels 20, evacuation channel 22, and otherchannels) and nano-wells 18, can be formed together with a base thatforms the bottom of each of the structures. For example, the base andstructure can be formed using any applicable method, for example, by amolding, spin coating, stamping process, hot embossing,three-dimensional (3D) printing, etc., or can be formed by applying anetching, micromachining, or photolithography process to a block ofmaterial. According to some embodiments a cover can then be attached tothe base and structure to cover the structure. Typically, the cover istransparent to enable optical or visual examination of the contents.Typically, the cover includes openings to enable introduction of liquidsinto the structure. For example, one or more openings can be positionedso as to enable introduction of liquids into common inlet 12, and, atleast in some cases, into one or more separate inlets 32. One or moreopenings 44 can be positioned to enable evacuation of air there-through,or application of negative pressure to evacuation channel 22.

According to some embodiments, the length (or, in some cases, thecross-sectional area, or both) of each distribution channel 24 isselected such that the rate of the flow of a liquid that is introducedinto that distribution channel 24, via common inlet 12, is substantiallyequal to the rate of flow in all of the other distribution channels 24.In the example shown, in order to achieve the equal flow rates, thelengths of each of distribution channels 24 b to 24 f is increased bythe addition of one or more extensions, such as open loops 26. In theexample shown, all open loops 26 are of substantially equal, havingpredetermined length, and are approximately U-shaped (e.g., with acurved or flat bottom). In the schematic example shown, the length ofeach open loop 26 is equal to separation distance d between two adjacentconnection nodes 40, where adjacent distribution channels 24 connect todistribution trunk channel 28. The number of open loops 26 added to eachdistribution channel 24 is selected to retard the rate of flow in adistribution channel 24 (e.g., in distribution channel 24 f) thatconnects common inlet 12 to a more proximal (e.g., to common inlet 12 orto inlet connection 36) SNDA component 14 to equal the rate of flow in adistribution channel 24 (e.g., distribution channel 24 a) that connectscommon inlet 12 to a more distal SNDA component 14.

It may be noted that, in the schematic example shown, the number of openloops 26 that are added to each distribution channel 24 is based on asimple calculation, in which the number of open loops 26 of length dthat are added to each distribution channel 24 b to 24 f that branchesoff of distribution trunk channel 28, at a connection node 40, is equalto the distance between that connection node 40 and the most distal node(e.g., the connection node 40, where distribution channel 24 a connectsto distribution trunk channel 28). A more accurate calculation thattakes into account different flow rates through different sections ofdistribution trunk channel 28 is described below.

In other examples, the lengths of different distribution channels 24 canbe otherwise adjusted, cross sectional areas of different distributionchannels 24 can be adjusted, surface properties of differentdistribution channels 24, or other adjustments to distribution channels24 can be made to achieve equal rates of flow through all distributionchannels 24.

According to some embodiments, when a pressure difference between commoninlet 12 and evacuation channel 22 is constant (e.g., due to negativepressure that is applied to evacuation channel 22), the rate of flow ineach distribution channel 24 of a liquid that is introduced intomultiplexed SNDA device 10, via common inlet 12, can be inverselyproportional to the resistance of each distribution channel 24 to flow(e.g., analogous to Ohm's law that states that current is equal topotential difference divided by electrical resistance). In the case oflaminar flow, resistance to flow can be a function of at least theviscosity of the liquid, cross sectional area of a conduit, and lengthof the conduit.

In the example shown, the cross-sectional areas of all distributionchannels 24, as well as of distribution trunk channel 28, aresubstantially identical. Therefore, in the event of laminar flow of asingle incompressible liquid through all distribution channels 24, therate of flow through a distribution channel 24 can be adjusted byadjusting the length of that distribution channel 24. Furthermore, itmay be assumed that the resistances to flow through all SNDA component14 of multiplexed SNDA device 10 are substantially identical. Therefore,it may be assumed that, when substantially equal flow rates areachieved, the difference in pressure between inlet connection 36 betweencommon inlet 12 and distribution trunk channel 28, and the connection(along SNDA device connection line 34) of each distribution channel 24to its connected SNDA components 14 is the same for all distributionchannels 24.

Accordingly, a calculation of a length of each distribution channel 24,or, equivalently, of a number of open loops 26 (of predetermined length)that are to be included in each distribution channel 24, can be based onan analogy to Kirchhoff's rules for electrical circuits.

According to some embodiments, in such an analogous calculation, thepressure difference between two points that are connected by one or moreconduits is analogous to a difference in electrical potential, orvoltage. As in the electrical analog, the pressure difference is thesame for all parallel conduits that connect the two points. The flowrate is analogous to electrical current. As in the electrical analog, ata node where a single conduit branches into two or more branch conduits,the total flow rate into the node (e.g., through the single node) isequal to the total flow rate out of the node (e.g., through all thebranch conduits). Resistance to flow in each conduit is analogous toelectrical resistance. Thus, as in Ohm's law of the electrical analog,the rate of flow in a conduit is equal to the pressure differencebetween the ends of the conduit divided by the resistance to flow inthat conduit.

Therefore, as in the electrical analog, when conduits are connected inseries, the total resistance to flow R_(s) is the sum of the resistancesto flow of the connected conduits:

R _(s) =R ₁ +R ₂ + . . . +R _(n),

where R₁, R₂, . . . R_(n) are the resistances to flow of each of theconnected conduits. Similarly, when n conduits are connected inparallel, the total resistance to flow R_(p) may be calculated from theformula:

1/R _(p)=1/R ₁+1/R ₂+ . . . +1/R _(n).

In an example where laminar flow may be assumed (e.g., slow flow ratesand low Reynold's number), and where all of the conduits have similarwalls and cross sections, the resistance to flow is substantiallyproportional to the length of the conduit. Therefore, in such a case,lengths of conduit sections may be substituted for the resistances inthe above formulae.

Multiplexed SNDA device 10 is configured to enable substantially equalflow rates through all of distribution channels 24. In particular,calculations based on the analogy to electrical current can be appliedto distribution trunk channel 28 and distribution channels 24 betweeninlet connection 36 and SNDA device connection line 34. The purpose ofthe calculation is to determine any additional resistance to flow thatis to be added to distribution channels 24, in order to enablesubstantially equal flow rates in all distribution channels 24.

According to some embodiments, by making the flow rates equal in alldistribution channels 24, all SNDA components 14 can be filledconcurrently and the terms applied on SNDAs are identical. In theabsence of a configuration that enables equal flow rates, an SNDAcomponent 14 that is nearest to common inlet 12 (e.g., an SNDA component14 that is connected to distribution channel 24 f) would be likely tocompletely fill before an SNDA component 14 that is further from commoninlet 12 (e.g., an SNDA component 14 that is connected to any ofdistribution channels 24 a to 24 e) has completed filling, or perhapshas not even begun to fill. Such uneven filling could adversely affectresults of testing that entails comparison of results in different SNDAcomponents 14 of multiplexed SNDA device 10.

FIG. 2 schematically illustrates an arrangement of distribution channelsof a portion of a multiplexed array of SNDA components, according tosome embodiments of the invention.

As shown in FIG. 2, all unlengthened distribution channels 42 a to 42 fare shown without any loops. As shown, unlengthened distributionchannels 42 a to 42 f are shown with their minimum lengths forconnecting inlet connection 36 with SNDA components 14, prior toadjustment in order to provide a uniform flow rate in all ofunlengthened distribution channels 42 a to 42 f. The length of each ofunlengthened distribution channels 42 a to 42 f, e.g., from itsconnection to distribution trunk channel 28 at one of connection nodes40 a to 40 f, to its connection to an SNDA component 14, at SNDA deviceconnection line 34, is channel minimum length D. The lateralcenter-to-center distance between adjacent connection nodes 40 a to 40 fis separation distance d.

In this example, since the path between inlet connection 36 and SNDAdevice connection line 34, via unlengthened distribution channel 42 a,is longer than the path via other unlengthened distribution channels 42b-42 f, any adjustments to the lengths of distribution channels 24 a to24 f may require lengthening of unlengthened distribution channels 42 bto 42 f, rather than shortening unlengthened distribution channel 42 a.In other examples/embodiments, e.g., where diagonal or other variants ofdistribution channels are allowed, adjustment can include shorteningdistribution channels.

According to some embodiments, the calculation yields a total channellength L, for each of distribution channels 24 a to 24 f, that enables auniform flow rate through all of the distribution channels 24 a-24 f. Asstated above, in the current example, total length L_(a) of distributionchannel 24 a between connection node 40 a and SNDA device connectionline 34 is equal to minimum length D.

According to some embodiments, at connection node 40 b, in order thatthe flow rate via distribution channel 24 b between connection node 40 band SNDA device connection line 34 equal that via distribution channel24 a, the resistances to flow via distribution channels 24 a and 24 b,and thus total lengths L_(a) and L_(b), respectively, are to be madeequal. The length of a path between connection node 40 b and SNDA deviceconnection line 34 via unlengthened distribution channel 42 a is the sumof D, the length of unlengthened distribution channel 42 a, and d, thedistance between connection node 40 b and connection node 40 a.Therefore, total channel length L_(b) for distribution channel 24 b(corresponding to unlengthened distribution channel 42 b, with an addedopen loop 26) can be calculated as:

L _(b) =D+d.

Accordingly, distribution channel 24 b includes an open loop 26 oflength d (or a plurality of loops whose total length is d).

According to some embodiments, at connection node 40 c, a calculatedtotal length L_(c) of distribution channel 24 c is to result in equalflow rates between connection node 40 c and SNDA device connection line34 via each of distribution channels 24 a to 24 c. Using theaforementioned formula for series and parallel resistances, theequivalent resistance to flow between connection node 40 c and SNDAdevice connection line 34 via parallel flow through distributionchannels 24 a and 24 b is proportional to (D+3d)/2. Further noting thatthe flow rate via the section of distribution trunk channel 28 betweenconnection node 40 c and 40 b (and thus through the combination ofdistribution channels 24 a and 24 b) is double the flow rate throughdistribution channel 24 c, the total length L_(c) of distributionchannel 24 c that enables a uniform flow rate can be calculated to be:

L _(c) =D+3d.

Accordingly, distribution channel 24 c includes one or more open loops26 of total length 3d. It may be noted that the length of open loops 26that are added to distribution channel 24 c in this calculation forL_(c) of distribution channel 24 c, as well as the calculations belowfor distribution channels 24 d to 24 f, differs from the number of openloops 26 shown in the general layout illustration in FIG. 1B, and whichare based on a different calculation.

Similarly, according to some embodiments, at connection node 40 d, acalculated total length L_(d) of distribution channel 24 d is to resultin equal flow rates between connection node 40 d and SNDA deviceconnection line 34 via each of distribution channels 24 a to 24 d. Usingthe aforementioned formula for series and parallel resistances, theequivalent resistance to flow between connection node 40 d and SNDAdevice connection line 34 via parallel flow through distributionchannels 24 a through 24 c is proportional to (D+3d)/3. Further notingthat the flow rate via the section of distribution trunk channel 28between connection node 40 d and 40 c (and thus through the combinationof distribution channels 24 a to 24 c is triple the flow rate throughdistribution channel 24 d, the total length L_(d) of distributionchannel 24 d that enables a uniform flow rate can be calculated to be:

L _(d) =D+6d.

Accordingly, distribution channel 24 d includes one or more open loops26 of total length 6d.

Similarly, according to some embodiments, at connection node 40 e, acalculated total length L_(e) of distribution channel 24 e is to resultin equal flow rates between connection node 40 e and SNDA deviceconnection line 34 via each of distribution channels 24 a to 24 e. Usingthe aforementioned formula for series and parallel resistances, theequivalent resistance to flow between connection node 40 e and SNDAdevice connection line 34 via parallel flow through distributionchannels 24 a through 24 d is proportional to (D+6d)/4. Further notingthat the flow rate via the section of distribution trunk channel 28between connection node 40 e and 40 d (and thus through the combinationof distribution channels 24 a to 24 d is quadruple the flow rate throughdistribution channel 24 e, the total length L_(e) of distributionchannel 24 e that enables a uniform flow rate can be calculated to be:

L _(e) =D+10d.

Accordingly, distribution channel 24 e includes one or more open loops26 of total length 10 d.

Finally (in the example shown), according to some embodiments, atconnection node 40 f, a calculated total length L_(f) of distributionchannel 24 f is to result in equal flow rates between connection node 40f and SNDA device connection line 34 via each of distribution channels24 a to 24 f. Using the aforementioned formula for series and parallelresistances, the equivalent resistance to flow between connection node40 f and SNDA device connection line 34 via parallel flow throughdistribution channels 24 a through 24 e is proportional to (D+10d)/5.Further noting that the flow rate via the section of distribution trunkchannel 28 between connection node 40 f and 40 e (and thus through thecombination of distribution channels 24 a to 24 e is five times the flowrate through distribution channel 24 f, the total length L_(f) ofdistribution channel 24 f that enables a uniform flow rate can becalculated to be:

L _(f) =D+15d.

Accordingly, distribution channel 24 f includes one or more open loops26 of total length 15 d.

According to some embodiments, this calculation can be continued in asimilar manner for numbers of distribution channels 24 greater than six.When the number of distribution channels 24 is fewer than six, thecalculation can proceed as described above until the lengths L of alldistribution channels 24 have been calculated.

It may be noted that, when distribution channels 24 are arrangedsymmetrically about symmetry axis 30, calculations need be performedonly on one side of symmetry axis 30. When symmetrically arranged, thecalculated total lengths L of each pair of symmetrically arrangeddistribution channels 24 that are equidistant from symmetry axis 30 areidentical to one another. In the event of an asymmetric arrangement ofdistribution channels 24, or where the distance between adjacentconnection nodes 40 is not the same for all pairs of adjacentdistribution channels 24, calculation may be modified in accordance withthe asymmetric positions of distribution channels 24.

FIG. 3 schematically illustrates distribution channels of the right sideof the symmetry plane of a multiplexed array of SNDA components,according to some embodiments of the invention, where the lengths of thechannels being adjusted to enable a uniform flow rate.

According to some embodiments, in channel arrangement 46, a total lengthof each of distribution channels 24 a to 24 d is as calculated in theexamples above. The length of each of distribution channels 24 b to 24 dincludes one or more open loops 26. In the example shown, the length ofeach open loop 26 is equal to separation distance d. Therefore, thenumber of open loops 26 in each of distribution channels 24 a to 24 d isequal to the multiple of d that is added to channel minimum length D toyield total length L for each of distribution channels 24 a to 24 d.

For example, in accordance with the calculation above, distributionchannel 24 a includes no (zero) open loops 26, distribution channel 24 bincludes one open loop 26, distribution channel 24 c includes three openloops 26, and distribution channel 24 d includes six open loops 26.Identical numbers of open loops 26 can be included in distributionchannels 24 that extend from distribution trunk channel 28 at positionsthat are symmetrical about symmetry axis 30 to those of distributionchannels 24 a to 24 d.

It may be noted that a maximum distance between distribution trunkchannel 28 and SNDA device connection line 34 can be limited by variousconsiderations. Accordingly, there can be various reasons for limitingthe number of open loops 26 that can be added to a distribution channel24. Other considerations can limit a minimum size of d. Thus, the numberof distribution channels 24 that extend from distribution trunk channel28 may be limited. In the examples shown in FIGS. 1B and 3, the maximumnumber of open loops 26 that can be included in a single distributionchannel 24 is limited to about six. In this case, if the added length iscalculated as described above, no more than four distribution channels24 can extend from distribution trunk channel 28 on either side ofsymmetry axis 30.

Alternatively, or in addition to adjusting a total length of eachdistribution channel 24, a cross section of each distribution channel 24can be designed to enable substantially identical flow rates through alldistribution channels 24. For example, channel arrangement in such acase can be similar to the arrangement of FIG. 2, where eachunlengthened distribution channel 42 has a different cross section.

For example, results of a flow simulation may yield a width of eachunlengthened distribution channel 42 required to provide identical flowrates through all of unlengthened distribution channels 42.

In one example simulation, the widths of unlengthened distributionchannel 42 a and of distribution trunk channel 28 were set to 150 μm(e.g., to match the width of primary channels 16), d was set to 2.35 mm,and D was set to 11 mm. In this simulation, the calculated widths rangedfrom 14 μm n for unlengthened distribution channel 42 b to about 10 μm nfor unlengthened distribution channel 42 f. It may be noted that, inthis example, the differences in width among unlengthened distributionchannels 42 b to 42 f are small relative to the width of unlengtheneddistribution channel 42 a. Different results can be obtained fromsimulations based on other dimensions.

According to some embodiments, the rectangular shape of multiplexed SNDAdevice 10 can enable connecting a plurality of component multiplexedSNDA devices 10 into a multi-array system. The multi-array system caninclude a single inlet port into which a liquid is to be introduced toflow to all the component multiplexed SNDA devices 10 via an arrangementof feeder channels. Similarly, all secondary channels 20 can beconnected to a single evacuation channel (e.g., having a rectangularform) to which negative pressure can be applied.

FIG. 4A schematically illustrates an example of channels of a system 51of multiple multiplexed SNDA devices, according to some embodiments ofthe invention, where all SNDA devices 10 a-10 h are oriented parallel toone another.

In the example shown of channeling system 50, eight multiplexed SNDAdevices 10 a-10 h, and their associated channel arrangements 46, areconnected to a single input port 52. A liquid that is introduced intochanneling system 50 via input port 52 can flow from input port 52 tomultiple channel arrangements 46 via feeder channels 54. Feeder channels54 are configured such that the lengths of all paths from input port 54to each of channel arrangements 46 are substantially identical. In theexample shown, feeder channels 54 are arranged in a branched pattern inwhich all branches are of equal length.

According to some embodiments, a single evacuation channel (not shown),for example having a rectangular shape or a U-shape, can surround all ofthe multiplexed SNDA devices 10 a-10 h that are connected to input port52, via feeder channels 54 and channel arrangements 46. According tosome embodiments, the evacuation channel can include a single port viawhich negative pressure can be applied to all component multiplexed SNDAdevices 10 a-10 h.

FIG. 4B schematically illustrates an example of channels of a system 61of multiple multiplexed SNDA devices 10 i-101, according to someembodiments of the invention, where some SNDA devices are orientedperpendicularly to others.

In the example shown of channeling system 60, four multiplexed SNDAdevices 10 i-101, and their associated channel arrangements 46 a and 46b, are connected to a single input port 52. A liquid that is introducedinto channeling system 60 via input port 52 can flow from input port 52to multiple channel arrangements 46 a and 46 b via feeder channels 62.In the example shown, feeder channels 62 are in the form of segmentswith resistance that can be substantially lower than the resistance at46 a and 46 b entry port, ensuring that all feeding channels are filledprior to reaching the 46 a,b complexes.

In the example shown, channel arrangements 46 a are arranged oppositeone another across input port 52. Similarly, channel arrangements 46 b,each rotated 90° to channel arrangements 46 a, are arranged opposite oneanother across input port 52.

According to some embodiments, a single evacuation channel (not shown),e.g., that is rectangular, can surround all of the multiplexed SNDAdevices 10 i-101 that are connected to input port 52 via feeder channels54 and channel arrangements 46 a and 46 b. The evacuation channel caninclude a single port via which negative pressure can be applied to allcomponent multiplexed SNDA devices 10 i-101.

Different embodiments are disclosed herein. Features of certainembodiments may be combined with features of other embodiments; thus,certain embodiments may be combinations of features of multipleembodiments. The foregoing description of the embodiments of theinvention has been presented for the purposes of illustration anddescription. It is not intended to be exhaustive or to limit theinvention to the precise form disclosed. It should be appreciated bypersons skilled in the art that many modifications, variations,substitutions, changes, and equivalents are possible in light of theabove teaching. It is, therefore, to be understood that the appendedclaims are intended to cover all such modifications and changes as fallwithin the true spirit of the invention.

While certain features of the invention have been illustrated anddescribed herein, many modifications, substitutions, changes, andequivalents will now occur to those of ordinary skill in the art. It is,therefore, to be understood that the appended claims are intended tocover all such modifications and changes as fall within the true spiritof the invention.

1. A device comprising: plurality of Stationary Nanoliter Droplet Array(SNDA) components (14); each SNDA component comprising: at least oneprimary channel (16); at least one secondary channel (20); and aplurality of nano-wells (18) that are each open to the primary channeland are each connected by one or more vents to the secondary channel;the vents are configured to enable passage of air solely from thenano-wells to the secondary channel, such that when a liquid isintroduced into the primary channel it fills the nano-wells, and theoriginally accommodated air is evacuated via the vents and the secondarychannel/s; wherein the plurality of the SNDA components are alignedparallel to one another and laterally displaced relative to one another,such that the device comprises a rectangular form; a single inlet port(12) and a distribution trunk channel (25,28) configured to enable asimultaneous introduction of the liquid into all primary channels; and asingle outlet port (44) and an evacuation channel (22) configured toenable a simultaneous evacuation of the air out of all the secondarychannels.
 2. The device of claim 1, wherein the diameter D_(DCh) or thesmaller side h_(DCh) of the distribution trunk channel (25) is selectedto be substantially larger than the diameter D_(PCh) or the smaller sideh_(PCh) of the primary channel/s, respectively D_(DCh)>D_(PCh) orh_(DCh)>h_(PCh); the distribution trunk channel is configured to befilled via the inlet port with liquid, while withholding the liquid fromthe primary channels, to about a predetermined threshold of its volume,enabling a liquid pressure formed there-within, to then simultaneouslyload all the primary channels.
 3. The device of claim 1, furthercomprising a plurality of distribution channels (24), each distributionchannel of the plurality of distribution channels connecting the inletport to the primary channel of a separate SNDA component; and whereineach distribution channel branches off from the single distributiontrunk channel (28) that is connected to the inlet.
 4. The device ofclaim 3, wherein each distribution channel (24) branches offperpendicularly from the distribution trunk channel (28).
 5. The deviceof claim 4, wherein each of the distribution channels (24) comprises adifferent cross section, relative to its distance from the single inlet(12), configured to allow a liquid to flow from the single inletopening, via the distribution trunk channel (28), and reach all of theSNDA components concurrently.
 6. The device of claim 4, wherein thedistribution channels (24) are arranged along the distribution trunkchannel (28) symmetrically, about a connection of the inlet to thedistribution trunk channel.
 7. The device of claim 4, wherein theconnections the plurality of distribution channels (24) with thedistribution trunk channel (28) are equally spaced along thedistribution trunk channel.
 8. The device of claim 4, wherein a totallength of each of each distribution channel of the plurality ofdistribution channels (24), between its connection to the distributiontrunk channel (28) and its connection to the primary channel (16) of anSNDA component (14), is adjusted to enable the substantially equal ratesof liquid flow.
 9. The device of claim 8, wherein the total length of atleast one distribution channel of the plurality of distribution channelsis lengthened by addition of one or more open loops (26) to said atleast one distribution channel (24).
 10. The device of claim 9, whereinthe lengths of all of the open loops that are added to distributionchannels of the plurality of distribution channels are substantiallyequal.
 11. The device of claim 10, wherein the length of an open loop ofsaid one or more open loops is equal to a distance between connectionsof two adjacent distribution channels of the plurality of distributionchannels to the distribution trunk channel, where the connections of theplurality of distribution channels to the distribution trunk channel areequally spaced along the distribution trunk channel.
 12. The device ofclaim 10, wherein the number of the open loops that are added to a firstdistribution channel is smaller than the number of the open loops thatare added to a second distribution channel, wherein a connection of thesecond distribution channel to the distribution trunk channel is moreproximal to a connection of the inlet to the distribution trunk channelthan to the connection of the first distribution channel to thedistribution trunk channel.
 13. The device of claim 3, wherein a crosssection of a distribution channel, of the plurality of distributionchannels (24), is selected to enable the substantially equal rates ofliquid flow into each of the primary channels (16).
 14. The device ofclaim 13, wherein a width of a distribution channel having a largestcross-sectional area, is equal to a width of the primary channel of theSNDA component to which that distribution channel is connected.
 15. Thedevice of claim 1, wherein all of the SNDA components are substantiallyidentical.
 16. The device of claim 1, further comprising a pressuredevice in communication with the outlet poet, configured to applysimultaneous negative pressure to all the secondary channels via theevacuation channel.